Rapid, on site, diagnosis of FMD and safe and cost-effective shipment of samples using lateral flow devices for laboratory diagnostics (A. Romey)
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The 2018 Open Session of the EuFMD Standing Technical Committee was held in Borgo Egnazia - Italy, 29-31 October 2018 . The session theme was on global vaccine security The European Commission for the Control of Foot-and-Mouth Disease (EuFMD), one of FAO’s oldest Commissions, came into being on the 12th June 1954, with the pledge of the sixth founding member state to the principles of a coordinated and common action against Foot-and-mouth Disease.
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Rapid, on site, diagnosis of FMD and safe and cost-effective shipment of samples using lateral flow devices for laboratory diagnostics (A. Romey)
- 1. RAPID, ON SITE, DIAGNOSIS OF FMD AND SAFE AND COST-EFFECTIVE SHIPMENT OF SAMPLES USING LATERAL FLOW DEVICES FOR LABORATORY DIAGNOSTICS Bakkali Kassimi L1, Belsham, G.J.2, Bulut A.N.3, Gorna, K.1, Hamers, C.4, Hudelet, P.4, Jamal, S.5, Laloy, E.1, Relmy, A.1, Romey, A.1, Ularamu H.G.6, Zientara, S.1 and Blaise-Boisseau S.1 1Anses, France, 2DTU, Denmark, 3SAP/FMD Institute, Turkey, 4Boehringer Ingelheim, France, 5University of Malakand, Pakistan, 6 NVRI, Nigeria Global Vaccine Security
- 2. Towards global control and eradication of FMD https://www.researchgate.net/publication/305238243_Genomics_and_outbreaks_Foot_and_mouth_disease; April, 2016. Global Vaccine Security - OS18: Oct 2018, Italy
- 3. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 4. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 5. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 6. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 7. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 8. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Use of lateral flow device for safe and low cost shipment of FMDV suspected samples (FMDVINACT)
- 9. Towards global control and eradication of FMD Global Vaccine Security - OS18: Oct 2018, Italy Use of lateral flow device for safe and low cost shipment of FMDV suspected samples (FMDVINACT)
- 10. Experimental protocol Global Vaccine Security - OS18: Oct 2018, Italy Viral suspension Elution Inoculation on cells 30min Soaking 15min in 0.2% C6H8O7
- 11. Experimental protocol Global Vaccine Security - OS18: Oct 2018, Italy Viral suspension RNA extraction Elution Inoculation on cells RT-rtPCR 3D & IRES rtPCR VP1 & sequencing 30min Soaking 15min in 0.2% C6H8O7 3DL VP4 VP2 VP3 2A 2B 2C 3A 3B 3CVP1 VPg5’ 3’ IRES 3B 3B Structural proteins Non-structural proteins
- 12. Experimental protocol Global Vaccine Security - OS18: Oct 2018, Italy Viral suspension RNA extraction Elution Inoculation on cells RT-rtPCR 3D & IRES rtPCR VP1 & sequencing Chemical transfection Virus rescue 30min Soaking 15min in 0.2% C6H8O7 3DL VP4 VP2 VP3 2A 2B 2C 3A 3B 3CVP1 VPg5’ 3’ IRES 3B 3B Structural proteins Non-structural proteins
- 13. Experimental protocol Global Vaccine Security - OS18: Oct 2018, Italy Viral suspension RNA extraction Elution Inoculation on cells RT-rtPCR 3D & IRES rtPCR VP1 & sequencing Chemical transfection Virus rescue 30min Soaking 15min in 0.2% C6H8O7 3DL VP4 VP2 VP3 2A 2B 2C 3A 3B 3CVP1 VPg5’ 3’ IRES 3B 3B Structural proteins Non-structural proteins • 7 reference strains • 3 archival samples
- 14. Field evaluation o Anses, France (coordinator) o Technical University of Denmark (DTU) o FMD Research Centre of Nigeria (NRVI) o FMD Institute of Turkey (SAP) o University of Malakand in Pakistan (UM) o Merial- Boehringer Ingelheim (BI) Evaluation in field conditions of a safe and cost-effective protocol for shipment of samples from FMD suspected cases for laboratory diagnostic (FIELD_EVAL_INACT) Global Vaccine Security - OS18: Oct 2018, Italy
- 15. Discard citric acid In farm Global Vaccine Security - OS18: Oct 2018, Italy Field evaluation on freshly collected clinical samples
- 16. Decontamination solution S1 S2 S3 In decontamination area Discard citric acid In farm Global Vaccine Security - OS18: Oct 2018, Italy Field evaluation on freshly collected clinical samples
- 17. Decontamination solution S1 S2 S3 In decontamination area Discard citric acid Triple packaging At room T S1 S2 S3 In clean area In farm Global Vaccine Security - OS18: Oct 2018, Italy Field evaluation on freshly collected clinical samples
- 18. Field evaluation on freshly collected clinical samples Global Vaccine Security - OS18: Oct 2018, Italy
- 19. Samples collection and inactivation of LFD in the field Global Vaccine Security - OS18: Oct 2018, Italy Turkey, n=20, completed Nigeria, n=20, completed Pakistan, n=400, in progress
- 20. Samples collection and inactivation of LFD in the field Global Vaccine Security - OS18: Oct 2018, Italy Turkey, n=20, completed Nigeria, n=20, completed Pakistan, n=400, in progress Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 21. Field evaluation on freshly collected clinical samples Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018Ularamu Hussaini, Nigeria, 2018 Naci Bulut, Turkey, 2018 Ularamu Hussaini, Nigeria, 2018 Naci Bulut, Turkey, 2018
- 22. Safety tests in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Nigeria Turkey
- 23. Safety tests in the lab 9Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Nigeria Turkey … in progress Elution Inoculation on cells 15min in 0.2% C6H8O7
- 24. Molecular detections in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Netherlands
- 25. Molecular detections in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Netherlands … in progress 15min in 0.2% C6H8O7 RNA extraction RT-rtPCR 3D & IRES rtPCR VP1 & sequencing
- 26. Virus rescue in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Netherlands
- 27. Virus rescue in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Netherlands … in progress RNA extraction 15min in 0.2% C6H8O7 Chemical transfection
- 28. Virus rescue in the lab Global Vaccine Security - OS18: Oct 2018, Italy Denmark France Netherlands … in progress RNA extraction 15min in 0.2% C6H8O7 Chemical transfection Optimization step
- 29. Optimization of RNA chemical transfection Global Vaccine Security - OS18: Oct 2018, Italy Chemical Reagents RNA/ transfectants ratios CPE on cells 24 hpt IBRS-2 ZZ-R-127 BHK-21 Lipo 2000 1 + ++ - 2 ++ +++ - 3 ++ +++ - 4 ++ +++ - 5 - + - 6 - ++ - 7 - ++ - 8 - ++ - Lipo 3000 1 - + - 2 +/- ++ - 3 + ++ - 4 + ++ - 5 - +/- - 6 - + - 7 - +/- - 8 - +/- - Messenger MAX 1 +++ ++ - 2 +++ ++ - 3 +++ +++ - 4 Toxicity +++ - 5 - ++ - 6 - ++ - 7 - ++ - 8 - ++ - Trans-IT 1 +++ ++ +/- 2 +++ ++ +/- 3 +++ + +/- 4 +++ + +/- 5 +/- +/- - 6 +/- +/- - 7 +/- +/- - 8 +/- +/- - Transfast 1 ++ ++ +/- 2 ++ ++ +/- 3 ++ ++ +/- 4 +/- Toxicity - 5 +/- Toxicity - 6 +/- Toxicity - Optimization step - 5 transfection reagents (Biswal et al., 2015; Bisht et al., 2014…) - 8 conditions per reagent - 3 cells lines
- 30. Optimization of RNA chemical transfection Global Vaccine Security - OS18: Oct 2018, Italy Chemical Reagents RNA/ transfectants ratios CPE on cells 24 hpt IBRS-2 ZZ-R-127 BHK-21 Lipo 2000 1 + ++ - 2 ++ +++ - 3 ++ +++ - 4 ++ +++ - 5 - + - 6 - ++ - 7 - ++ - 8 - ++ - Lipo 3000 1 - + - 2 +/- ++ - 3 + ++ - 4 + ++ - 5 - +/- - 6 - + - 7 - +/- - 8 - +/- - Messenger MAX 1 +++ ++ - 2 +++ ++ - 3 +++ +++ - 4 Toxicity +++ - 5 - ++ - 6 - ++ - 7 - ++ - 8 - ++ - Trans-IT 1 +++ ++ +/- 2 +++ ++ +/- 3 +++ + +/- 4 +++ + +/- 5 +/- +/- - 6 +/- +/- - 7 +/- +/- - 8 +/- +/- - Transfast 1 ++ ++ +/- 2 ++ ++ +/- 3 ++ ++ +/- 4 +/- Toxicity - 5 +/- Toxicity - 6 +/- Toxicity - MockCPE on ZZ-R-127 48h post-transfection
- 31. Optimization of RNA chemical transfection Global Vaccine Security - OS18: Oct 2018, Italy Chemical Reagents RNA/ transfectants ratios CPE on cells 24 hpt IBRS-2 ZZ-R-127 BHK-21 Lipo 2000 1 + ++ - 2 ++ +++ - 3 ++ +++ - 4 ++ +++ - 5 - + - 6 - ++ - 7 - ++ - 8 - ++ - Lipo 3000 1 - + - 2 +/- ++ - 3 + ++ - 4 + ++ - 5 - +/- - 6 - + - 7 - +/- - 8 - +/- - Messenger MAX 1 +++ ++ - 2 +++ ++ - 3 +++ +++ - 4 Toxicity +++ - 5 - ++ - 6 - ++ - 7 - ++ - 8 - ++ - Trans-IT 1 +++ ++ +/- 2 +++ ++ +/- 3 +++ + +/- 4 +++ + +/- 5 +/- +/- - 6 +/- +/- - 7 +/- +/- - 8 +/- +/- - Transfast 1 ++ ++ +/- 2 ++ ++ +/- 3 ++ ++ +/- 4 +/- Toxicity - 5 +/- Toxicity - 6 +/- Toxicity - MockCPE on ZZ-R-127 48h post-transfection 2 transfection reagents 1 condition per reagent 1 cell line … in progress
- 32. Conclusion Global Vaccine Security - OS18: Oct 2018, Italy Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 33. Conclusion Global Vaccine Security - OS18: Oct 2018, Italy Soaking 15min in 0.2% C6H8O7 Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 34. Conclusion Global Vaccine Security - OS18: Oct 2018, Italy Soaking 15min in 0.2% C6H8O7 Romey,Guinée,2018 Ularamu Hussaini, Nigeria, 2018 Ularamu Hussaini, Nigeria, 2018
- 35. Conclusion Global Vaccine Security - OS18: Oct 2018, Italy Soaking 15min in 0.2% C6H8O7 Effective vaccine selection UlaramuHussaini,Nigeria,2018 Romey,Guinée,2018 Ularamu Hussaini, Nigeria, 2018
- 36. Conclusion Global Vaccine Security - OS18: Oct 2018, Italy Soaking 15min in 0.2% C6H8O7 Effective vaccine selection UlaramuHussaini,Nigeria,2018 Romey,Guinée,2018 Ularamu Hussaini, Nigeria, 2018
- 37. Grazie a tutti per l’attenzione e per l'organizzazione